Α-Synuclein Exhibits Differential Membrane Perturbation, Nucleation, and TLR2 Binding Through Its Secondary Structure.

Amyloid formation drives the pathology of different neurodegenerative diseases. α-Synuclein is a natively unfolded protein that assembles itself into toxic amyloid structures, hence contributing to synucleinopathy. Its amyloid formation proceeds through various conformational intermediate stages, starting with a lag phase, followed by a rapid growth phase, and leading to beta rich fibril formation. Few studies have shown that the helix rich intermediate may be involved in fibril formation. Earlier, the helix intermediate was only studied in the membrane bound state. Despite many years of research, a precise mechanism of α-synuclein aggregation and the significance of intermediates with variable secondary structures are not well elucidated. Therefore, this study aims to understand the importance of secondary structures in α-synuclein-mediated neuronal toxicity. Our data revealed that the helix rich intermediate species exposes more of the hydrophobic surface than the beta rich intermediate species and harbors with the lipid membrane efficiently, thus contributing to the greater roughness of the cellular membrane that subsequently results in membrane disruption. It has been seen that upon internalization these species also activate the redox machinery. β-Sheet enrichment contributes to self-assemblies of monomeric α-synuclein as it binds more with the monomeric species than the helix rich species. Additionally, we also observed that the beta rich species exhibits stronger TLR2 binding than the helix rich species as well as a potentiated neuroinflammatory cascade. Taken together, our data evidently put forward that secondary structures play a differential role during amyloid formation, and targeting them can be a novel intervention strategy for neurodegenerative disease progression.